Purification and Optimization of Extracellular Lipase from a Novel Strain Kocuria flava Y4

Abstract

The exogenous lipolytic activities of Kocuria sp. have been recognized earlier but the genus further contains many more unexplored strains. In this study, the extracellular lipase activity of Kocuria flava Y4 (GenBank accession no. MT773277), isolated from Dioscorea villosa during our previous study, was regulated by different physicochemical parameters, such as pH, temperature, shaking speed, and incubation time. For efficient immobilization of the extracellular lipase, 4% sodium alginate, 50 mL of 25 nM CaCl2.2H2O solution, and 15 min. Hardening time of gel beads in calcium chloride was used. For the first time, K. flava Y4 lipase was purified using ammonium sulphate precipitation followed by dialysis and DEAE-Sepharose anion exchange chromatography with Sepharose-6B gel filtration chromatography, yielding ∼15-fold purified lipase with a final yield of 96 U/mL. The SDS-PAGE of purified lipase displayed a single strong band, indicating a monomeric protein of 45 kDa. At a temperature of 35°C and pH 8, the purified lipase showed maximum hydrolytic activity. Using p-nitrophenyl acetate (p-NPA) as the hydrolysis substrate, the values of Km and Vmax derived from the Lineweaver–Burk plot were 4.625 mM and 125 mol/min−1mg−1, respectively.

Optimization of Culture Conditions for Lipase Production by Pseudomonas aeruginosa ECS3

“…This indicates that Pseudomonas aeruginosa can produce lipase at 35 o C. It appears that the lipase activity slightly increased at 45 o C, possibly due to environmental factors (Figure 2). Similarly, lipase production by Pseudomonas sp., Penicillium fellutanum, Kocuria flava Y4, and Providencia stuartii was stimulated at 35 o C. [26][27][28][29] Lipase from Staphylococcus sp., Bacillus cereus, and Geobacillus stearothermophilus, on the other hand, were maximally obtained at 37 o C, 30 45 o C, 31 and 60 o C, 32 respectively. This shows that the physical properties of the cell membrane, energy metabolism, and translational synthesis of proteins are affected by temperature.…”

Deciphering the underlying core microorganisms and the marker compounds of Liupao tea during the pile‐fermentation process

“…Kocuria (the phylum Actinobacteria) was also found in PET during the fermentation stage 2 . The members of this genus have extracellular lipase activity, extracellular amylase, and laccase activity 20,21 . Kocuria koreensis , previously isolated in fermented seafood, was positive for catalase, esterase, esterase lipase, β ‐glucosidase, ans so on, 22 and we isolated it from LBT during the pile fermentation.…”

“…Correlation analysis showed that Staphylococcus , Kocuria , Blastobotry , and Trichomonascus were positively correlated with α ‐linolenic acid. Kocuria has extracellular lipase activity, 21 and Trichomonascus can secrete lipase 31 . However, the mechanism by which these microorganisms promote α ‐linolenic acid production needs to be studied further.…”

“…2 The members of this genus have extracellular lipase activity, extracellular amylase, and laccase activity. 20,21 Kocuria koreensis, previously isolated in fermented seafood, was positive for catalase, esterase, esterase lipase, ⊎-glucosidase, ans so on, 22 and we isolated it from LBT during the pile fermentation. Nevertheless, the role of K. koreensis in LBT is not well elucidated.…”

Evaluation of enzymatic degradation of ochratoxin A by protease and lipase produced by lactobacillus isolated from dairy products

“…Lipase and protease produced by microbial are more valuable compared to those derived from plants or animals due to their variety of catalytic activities available, high yield production, simplicity of genetic manipulation, absence of seasonal fluctuations, regular supply, more stability, and more safety and convenience, and the growth rate of microorganisms is very high in economical media. [23,24]. 1, and 37.03%, respectively.…”